Test Protocol for Legionella: Science Sunday

At our lab we are certified for two different methods for identifying and quantifying Legionella in samples. One is the general CDC method and the other is a method that is WAY more involved and only required by the State of New York.  Why the state of New York has to be extra special is beyond me, but we have been audited by them and found to do everything properly so we can do New York samples.

In the following post, I actually am doing both methods simultaneously because one client is from not New York, and one client is from New York. Many of the steps are similar. Here we go!

Here is nearly everything I need to test for both methods of Legionella. From left to right:  vacuum pump, 100ml sterilized tips, pipetter, erlenmeyer flask with rubber fitting, vortexer, sharpie, chlorine test strips, samples, skinny tubes, glass tubes, black agar plates, blue large tubes, Page's Saline, saline, NY acid, Legionella acid, cultures of Pseudomonas aeruginosa and Legionella pneumoniae, sterile water, 2um disposable filters.

Not pictured: 1 ml glass pipettes, pipette bulb, forceps, ethanol, lighter

First step, required for NY, optional for CDC.  Test the chlorine content of the water.

Good result! You can proceed with testing of the sample.

BAD RESULT.

Add sodium thiosulfate until result is good. In this case it was 100 ml of sample, so I added 100 ul at a time.

Fill large tubes with 10 ml of saline (Pages for NY, general for CDC)

Filter 100 ml of sample.

Dip the forceps into ethanol and SET THEM ON FIRE! (to sterilize) Once cool, use the forceps to transfer the filter paper to the blue eppendorf tubes.

VORTEX!  Just... generally for CDC, for 2 VERY LONG MINUTES for NY.

Yay!

Forceps and ethanol

For NY, you have to run positive and negative controls. Using the McFarland standards, you can guesstimate your concentration of bacteria in liquid by comparing the cloudiness.  Pseudomonas is the negative control, Legionella is duh, the positive.

To save time, vortex multiple tubes at once and cut vortexing time in half!

CDC:  Use bi plates with two similar agars on each side. Pipette 0.1 ml onto each side.

Spread that sample around. You can use the same spread from side 1 to 2, but not 2 to 1 and obviously not from one sample to the next.

NY samples go on a single type of agar (Side 2 but poured onto a whole plate). You plate 0.5 ml and spread.

Not pictured:  Each method has an acid treatment stage. This is pretty neat. The acid will kill amoebas that can hide Legionella. Yay! CDC has a 15 minute acid treatment.  NY has a 5 minute acid treatment in a different acid.

NY also has a 30 minute 50deg C heat treatment.

Each treatment also gets plated onto it's own plate.

In the end, CDC method uses 4 plates (regular and 3 acid) per sample, NY uses 3 plates per sample (regular, heat, acid) plus the controls.

All of those plates go into anaerobic jars.  You light the candle and tighten the lid, the candle burns all the oxygen. Those jars get incubated for 5 days (CDC) or up to 10 days (NY) at 35 deg C.

How we identify legionella coming up next!

BONUS SCIENCE

A picture of the product my company manufactures. It's a two sided dip stick. You dip it in a limited volume of water and bacteria or fungi stick to the agar. You can enumerate how much bacteria is in your volume of water.

Kid Pic Spam plus other stuff

An addendum to my previous post about Thinx, I'm also planning to try sea sponge tampons to see if they're more comfortable for riding (and pair with the thinx)  compared to my cup or disposable tampons.  The sponges are reusable for 3-6 months so you're still spending less and the product is somewhat renewable.

Anyway, I'll have the sponges in time for my next cycle, so I'll update then with how it all goes.


Hazel is making significant developmental progress. She can now easily crawl with her belly off the ground, is cruising around the house like a pro (when they walk but always hang on to stuff for balance) and has taken a single solo step on a few occassions.

She gets super  excited when she sees me pull out my carrier and loves being worn while I do stuff outside or go to the store. In general, she loves being the center of attention. Her favorite thing to say right now is Dada!  Which of course makes Eric very happy. I'm still getting her to work on Mama.

She loves Gwyn, Saffron and the cats and will screech excitedly/chase them.

She loves sitting in the high chair up above everyone. I think she likes watching them all.

Thinking about walking

We had a bout of sickness and she was diagnosed with RSV and had to have some breathing treatments. She's all better now though!

Sick baby passed out in the car. 

Using her toy to walk.

Two bottom teeth!

We did her 9 month photos a few days late.

Kaylee of course always wants to be in the pictures too.

Fun with colors at my job

The iridescent colors separate, it's really neat.

Kaylee insisted on riding in the snow.

Hazel is the center of attention while Kaylee is in gymnastics.

Kaylee is doing really well for her age group and she enjoys her once a week activity.

Eric is encumbered.

Side carred crib is also for cats!

A frequent sight.

Kisses for the kitty.

Sunrise on wednesday, my super busy day

I toured a school that I want Kaylee to attend for Kindergarten. It's a charter school that only accepts 40 students. Open enrollment ends Friday. This is the upper school (2nd through 8th grade)

Gym

Lower school (K-1)

Lower school recess. They are outside every day, regardless of weather.

Happy upside down baby.

Poking at Eric's moles.

Thinx Period Underwear

Content Warning:  Frank talk about menstruation, vaginas

Something New for 2017

This is a kind of horsey post, I'll likely xpost it to my horse blog too. 

Since moving to Michigan I've gotten pretty involved in the Great Distance Derby, which has connected me with some local, likeminded horse people. I've gone to Flint to donate water with them on two occassions and gone on some trail rides. I even 'walked' a trail ride with Gwyn in hand just to get out and socialize while I was pregnant since I wasn't allowed to ride. 

The person who organizes GDD has also started organizing a Michigan and Midwest Mounted Archery organization, and is bringing clinics to the area. I wasn't able to do anything last year, but I have done some mounted archery with the SCA back when I lived in Florida. I was a fan of it then and I see no reason why I could do it now.  At this point I'm trying to dial back my expectations of what competitions we can do without really sinking time and money into some training for Gwyn and myself and being realistic about what is within our reach and what is possible in the future.  

Anyway, that's musings for a whole post on its own. Suffice it to say, mounted archery is a pretty darn reasonable hobby and the clinics are reasonably local and kind of budget friendly with the added bonus of including camping! Yay! I love camping!

So for christmas, Eric got me a very basic, junior archery set.  Two arrows, a cheap bow and finger and arm guards that are not meant for my fingers or arms. 

A nice January day! 

For winter training, my friend Heather organized a group of us to meet up and do an activity called Archery Tag at a local indoor sports complex. 

You get totally cool face shields that fog up the moment you start sweating.

And play on an indoor field with fake grass and 'obstacles' 

It's basically like dodgeball but with a bow and arrow.

AND SO MUCH FUCKING FUN OMG

The arrows are blunted foam and while you can feel them make contact, you aren't hurt, mainly because each team is kept separate from one another. You don't cross into the other team's territory and the  neutral middle is where you aren't allowed to shoot from or be shot. 

You can see me in the top part of the picture aiming at Heather, lol.

I've just let loose an arrow!

The game starts with all the arrows in the middle, Hunger Games style like the Cornucopia. 

The referee counts down and then it's a mad dash to the middle to grab arrows and then back to be out of range (or at least at an easily dodgeable distance)  or hop behind an obstacle to snipe the other team.  

You get someone out by hitting them with an arrow. But, if you grab an arrow, the person who shot that arrow is out and someone on your own team can rejoin the game.

I like hiding behind things, but the constant crouching and standing up to shoot and dropping back down. Let's just say I did a LOT of squats. My legs burned FOR DAYS. 

You can also see the white target with black spots. The game was won if you either took out all the opposing team by hitting them or if you knocked out all five of those foam circles. Our team won boths ways!  

Team "Don't Shoot Me!"  which is retrospect was in poor taste....

The plan is to do this monthly in the cruddy winter months (Feb and March) before clinics start up and riding can begin in earnest again.  Until then, I'll be using my baby bow to desensitize Gwyn because I'm planning to do a clinic in June!  I'm so excited!

Year in Review

I haven't posted since the end of September. Damn.

Anyway, onward to a quick monthly recap because I can.

January
We made the most progress we'd make all year with our aquaponics project by acquiring cinderblocks and the pickle barrels.
I had lots of little rides on Gwyn despite doctors orders but felt MUCH better because of it all.
I got the nursery painted and Kaylee helped.

February
I had a very against doctors orders trail ride and felt spectacular
I tried to make Hazel a blanket and didn't get very far
We got a harness for skijoring and introduction for Gwyn pulling something behind her went... off the rails so to speak and Kaylee got upset that I broke her sled.
We started the process of putting in tile where we had hardwood!

March
I made a trip to Ikea all by myself while hugely pregnant and it was awesome
I got norovirus and dropped a ton of weight
I made monthly horse goals that I largely met!
We made good progress with the floors
We toured the hospital where I'd be giving birth.

April
We finished the floors and they still look fantastic.
I ended up in the ER again with breathing issues. No more clots but we finally figured out why I had a 4 month long cough (reflux)
Hazel finally flips head down and my huge worries about labor ease up
I anxiously wait through many bouts of false labor before finally being induced
Hazel is born!

May
I spent most of this month snuggling new baby
I also rode for the first time post baby

June
We did a test run of the water circulation for aquaponics
I had more baby snuggles
We enjoyed the start of summer and Hazel was a portable potato
I set up the electric fence tape so Gwyn and Saffron could assist in lawn mowing, much to Eric's delight

July
I find time to ride where I can with the assistance of baby monitors and long naps
We acquire an ATV and a small trailer to assist with farm chores
Gwyn learns to hi-tie
Kaylee turns 4
We visit a cool fossil park in Canada
I get a paid writing gig!

August
I start ramping up prep for endurance rides
Gwyn and I do the intro ride at the Metropark Express and easily meet our goals
I finally find a riding safety vest that fits me!
I get an interview for a microbiology job and get the job!

September
I start the new job and LOVE it, even if Hazel refusing to eat from a bottle is stressful
We explore the back roads and I find beautiful horse and hunt country finally
My posting starts to significantly drop off since I'm not at home

October
Busy horse month!
I compete in the Hunter Trials
I meet my friend Heather on my birthday for a Hunt Pace (which I never posted pictures of!)
I compete the 30 mile LD at Oak Leaf Run and camp with Gwyn solo for the first time!
Kaylee went trick or treating with her Aunt while I hung out around a fire with Hazel. Very pleasant

November
Not much going on. I rode maybe once.
Dealt with family drama over who was hosting Thanksgiving. Had a good holiday in the end

December
Decorated for Christmas and dealt with the arrival of the deep cold
Rode once, on New Years Eve
Family came to visit for Christmas, which was really awesome, my brother got to meet Hazel
Hazel starts crawling and cruising (walking around while hanging onto stuff)
Eric got a new car (Chevy Volt) and I'm using his old one (Sonic) so that my commute cost in gas can be reduced and so that I don't go over miles for the lease of the truck


So there you have it! You get a quick run down of the stuff I didn't blog about because I just didn't have time. We had a rough go of bedtime for a while so I couldn't escape Hazel and sit on the computer uninterrupted. And I just don't like blogging on my phone or tablet, which is what I'm using when I'm nursing Hazel.